1. BPDCN cells are dependent on BCL2 and sensitive to venetoclax in vitro.
BPDCN cells are dependent on BCL2 and sensitive to venetoclax in vitro. A, Representative photomicrograph of a bone marrow biopsy demonstrating involvement with BPDCN by hematoxylin and eosin (H&E) staining and BCL2 immunohistochemistry are shown. B, Western blotting in BPDCN (CAL1) and AML (KASUMI1, OCI-AML2, THP1, MOLM13, U937, and TF1) cell lines is shown for the indicated anti- and proapoptotic proteins. C, Baseline BH3 profiling is shown in the indicated cell lines. “Overall priming” represents cytochrome c release in response to BIM peptide stimulation at four concentrations (0.03 μmol/L, 0.1 μmol/L, 0.3 μmol/L, and 1 μmol/L); “BCL2/BCLXL dependence,” BAD peptide (10 μmol/L, 100 μmol/L); “BCLXL dependence,” HRK peptide (100 μmol/L); and “MCL1 dependence,” MS1 peptide (10 μmol/L). Graphs represent mean ± SEM of three independent experiments. D, Dynamic BH3 profiling depicting the difference in overall apoptotic priming before and after exposure to 100 nmol/L venetoclax for 4 hours (“delta priming” or “Δ% priming,” graphs represent mean ± SEM of three independent experiments; **, P < 0.01; ***, P < vs. CAL1 by t test). E, MTT assay results for the indicated cell lines after 72 hours of exposure to the indicated doses of venetoclax. Data points represent mean of three replicates ± SEM, line represents nonlinear regression (curve fit). F, Correlation of 4-hour dynamic BH3 profiling with 24-hour percent cell death by Annexin V/propidium iodide flow cytometry after exposure to 100 nmol/L venetoclax (Spearman r and P values for correlation are shown).
Joan Montero et al. Cancer Discov 2017;7:
©2017 by American Association for Cancer Research